We have obtain a steady and reliable dyeing methods for the uniuncleate and dicaryotic hyphae of Pleurotus tuber-regium by using different foster hyphae way, comparing two kinds of fastness liquid and three dye stuff on the hyphae nuclear stain effect, and then optimization grouping.

OBJECTIVETo monitor the change of renin, endothelin and 6-keto-prostaglandin F1alpha in canine during left ventricular assist (LVA).

METHODSEight canines were assisted by left assist ventricular device for 9 hours. The level of renin, endothelin and 6-keto-prostaglandin f1alpha in plasma were measured by radioimmunity analysis before assisting (control group) and at 3 hours, at 6 hours, at 9 hours after assisting.

RESULTSThe level of endothelin in plasma didn't dropped remarkably as LVA proceeded in which there was not differency in statistical diffency compared with control group[(51 +/- 11) ng/L, (42 +/- 8) ng/L, t = 0.926, P > 0.05]; The level of renin in plasma reached the summit at 3 hours during LVA compared with control group (3,036 +/- 1,411) ng/L, (1,783 +/- 467) ng/L, t = 5.013, P < 0.01) and later show dropping tendency without statistical differency at 9 hours (1 944 +/- 883) ng/L (t = 0.644, P > 0.05); The level of 6-keto-prostaglandin f1alpha in plasma at 3 hours during assisting increased remarkably [(75 +/- 17) ng/L, t = 1.411, P < 0.05), at 6 hours reached summit [(92 +/- 18)ng/L, t = 3.533, P < 0.01) and at 9 hours show dropping tendency with significant differency compared with control group (90 +/- 22) ng/L, t = 2.516, P < 0.05).

CONCLUSIONDuring 9 hours LVA, endothelin didn't dropped remarkably compared with control group and the endothelium released renin with transient increase, prostaglandin with consistent increase.

Animals ; Dogs ; Endothelins ; blood ; Heart-Assist Devices ; Models, Animal ; Prostaglandins ; blood ; Renin ; blood ; Ventricular Function

BACKGROUNDAllergic asthma is associated with airway inflammation and hyperresponsiveness caused by dysregulated production of cytokines secreted by allergen-specific helper T-type 2 (Th2) cells. The linker for activation of T cells (LAT) is a membrane-associated adaptor protein, which has been shown to take part in regulating T cell receptor (TCR) signaling and T cell homeostasis. In this study, we established an asthmatic mouse model to examine the changes in LAT levels during allergic airway disease and the effects of LAT transgenic expression on airway inflammation.

METHODST cells from mouse lung tissues were isolated from allergen challenged (ovalbumin (OVA)) and control mice, and the purity of these isolated T cells was examined by fluorescence-activated cell sorter (FACS). Semi-quantitative RT-PCR and Western blotting were used to detect the expression of the LAT gene and LAT protein, respectively. After an intranasally administered mixture of pCMV-HA-LAT plasmid and Lipofectamine 2000, 24 hours before and 72 hours after allergen challenge, the BALF cell count and the differential cytologies were studied. In addition, IL-4 and IFN-γ levels in the BALF were determined by ELISA, and pathological changes in lung tissues were observed.

RESULTSLAT protein and mRNA expression were decreased in lung T cells in a mouse model of allergen-induced airway disease. After intranasal administration of pCMV-HA-LAT, histopathological examination of the lungs showed that intervention with LAT overexpression prevented mice from developing airway inflammation, and the number of total cells, eosinophils, neutrophils, and lymphocytes in the BALF was reduced significantly compared with the OVA sensitized and challenged group. In addition, the Th2 cytokine IL-4 decreased, while the Th1 cytokine IFN-γ increased compared to the OVA sensitized and challenged group or the OVA sensitized group plus pCMV-HA treatment.

CONCLUSIONThis study demonstrates that LAT might effectively diminish Th2 cytokine responses, lung histopathological changes and lung inflammation to allergen challenge in a model of experimentally induced asthma.

Animals ; Asthma ; immunology ; metabolism ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; immunology ; Cells, Cultured ; Cytokines ; metabolism ; Female ; Inflammation ; immunology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; metabolism

OBJECTIVETo develop an inflow cannula of left ventricular assist implanted by blood vessel.

METHODSThe maximum inflow and properties against folding of 8 sorts of cannulae were measured in mimic extracorporeal circulation appliances and canines.

RESULTSThe maximum flow of the cannula increased, as the inner diameter became greater (P < 0.01) compared with each group. The maximum flow rate was (1.82 +/- 0.03) L/min, (2.44 +/- 0.03) L/min, (3.02 +/- 0.04) L/min, (3.31 +/- 0.03) L/min respectively for polyvinyl cannulae with wall thickness of 0.5 mm (PV 0.5 cannula) and inner diameter of 3 mm, 4 mm, 5 mm, 6 mm; (1.83 +/- 0.03) L/min, (3.07 +/- 0.04) L/min respectively for the polyvinyl chloride cannula with wall thickness of 1.0 mm imbedded by spring wire (PVCSW 1.0) and inner diameter of 3 mm and 5 mm; (1.82 +/- 0.02) L/min, 1.84 +/- 0.02 L/min for strengthened polyvinyl cannula with wall thickness of 0.8 mm (SPV 0.8) and inner diameter of 3 mm and polyvinyl cannula with wall thickness of 1.0 mm (PV 1.0 cannula) of inner diameter of 3 mm. There was no remarked statistical difference in vitro maximum flow among the four cannulae of 3 mm inner diameter in vitro. PVCSW 1.0 was showed the best antifolding property, PV 1.0 cannula good and SPV 0.8 and PV 0.5 unsatisfactory in properties against fold. There was no significant statistical difference between in vivo and in vitro maximum flow for PVCSW 1.0 and PV 1.0 cannulae of 3 mm inner diameter. But for SPV 0.8 and PV 0.5 cannulae of 3 mm inner diameter, there was a significant difference between in vivo and in vitro.

CONCLUSIONSPV 0.5 cannula and SPV 0.8 cannula are not suitable to clinical use. PV 1.0 cannula can be used in clinics. PVCSW 1.0 cannula is fully qualified for inflow conduit of left ventricular assist in surgery.

Animals ; Catheterization ; Dogs ; Heart-Assist Devices

OBJECTIVESlow transit constipation (STC) is a colonic motor disorder whose etiology remains unclear. Recent studies have demonstrated a crucial role for interstitial cells of Cajal (ICC) in regulation of intestinal motility. The aim of this study was to examine the distribution of ICC within the normal sigmoid colon and STC patients.

METHODSTwelve patients with STC and eight age-matched controls were studied. ICC were identified with a monoclonal antibody to c-kit by an indirect immunofluorescence method. Immunostained tissues were examined with a laser scanning confocal microscope and the area occupied by ICC was calculated with image analysis software.

RESULTSICC were located in the external muscle layers including longitudinal muscle (LM), myenteric plexus (MP), circular muscle (CM) and submucosal border (SMB). Two types of Kit-positive ICC were observed: bipolar cells characterized by one or two long processes, and multipolar cells with long stellate processes extending in various directions. A higher percentage of ICC was present in the MP regions and CM layers compared with the SMB and LM layers. Tissues from STC patients showed considerably decreased in number of ICC located in the four regions (ICC-LM, ICC-MP, ICC-CM, ICC-SMP), especially for ICC-SMP, almost completely disappeared.

CONCLUSIONDecreased c-kit + ICC in number may play an important role in the pathophysiology of STC. It remains to be determined whether loss of ICC is primary or secondary to another lesion.

Adult ; Aged ; Case-Control Studies ; Colon, Sigmoid ; pathology ; Constipation ; pathology ; physiopathology ; Female ; Fluorescent Antibody Technique, Indirect ; Gastrointestinal Transit ; physiology ; Humans ; Male ; Middle Aged

BACKGROUNDThe immunologic response to allergens mediated by T lymphocytes is an incipient key element in the pathogenesis of asthma, and Th1/Th2 balance is regarded as the core of asthma pathogenesis. Notch is a single-pass transmembrane receptor protein that regulates differentiation, proliferation and apoptosis in a broad range of cells. It is considered that the Notch signal pathway works in every stage of T cell development and differentiation. Whether the pathway of asthma pathogenesis is related to Notch1 remains unknown. This study is aimed to investigate whether the pathway of asthma pathogenesis is related to Notch1 by examining the effect of knockdown of the Notch1 gene by small interfering RNA on T cell differentiation.

METHODSAn OVA-induced asthma mouse model was established. The expression of Notch1 in the tissue and T cells of the lung from asthmatic mice was detected by RT-PCR and Western blotting. The expression of Notch1 and cytokine interleukin (IL)-4 and interferon (IFN)-gamma in activated lung T cells was detected by RT-PCR and enzyme-linked immunosorbent assay after blocking Notch1 by small interfering RNA.

RESULTSThe mRNA and protein expression of Notch1 increased significantly both in the lung tissue and lung T cells of asthmatic mice (both P < 0.05). IL-4 decreased and IFN-gamma increased significantly in active lung T cells after Notch1 was blocked by Notch1-specific small interfering RNA (IL-4: (2.51 +/- 0.51) pg/ml vs 0.64 +/- 0.27) pg/ml protein; IFN-gamma: (21.72 +/- 4.24) pg/ml vs (39.79 +/- 4.09) pg/ml protein, P < 0.05).

CONCLUSIONThis study demonstrated that the Notch1 signal might play a role in the pathogenesis of asthma by its involvement in Th1/Th2 differentiation.

Animals ; Blotting, Western ; Female ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lung ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; genetics ; metabolism ; Receptor, Notch1 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes ; metabolism

OBJECTIVETo determine the distributions of six Helicobacter pylori (Hp)-specific antibodies in a high-risk population of gastric cancer (GC) and explore the relationship between Hp virulence factors and precancerous gastric lesions.

METHODSBased on the two intervention trials conducted in Linqu County, the seropositivities for CagA, VacA, GroEL, UreA, HcpC and GGT were assessed by recombinant immunoassay (recomLine) in 623 participants with H. pylori infection determined by (13)C-urea breath test ((13)C-UBT) and/or enzyme linked immunosorbent assay (ELISA).

RESULTSIn a total of 623 participants were detected by recomLine analysis, of which 594 were Hp-positive. The seropositivities rates of CagA, VacA, GroEL, UreA, HcpC and GGT were 84.0%, 38.2%, 66.7%, 17.7%, 58.8% and 42.8%, respectively. A total of 523 participants were determined as type I infection of Hp, accounting for 88.1%. Compared with superficial gastritis (SG), the infection rate of Hp type I was higher in the chronic atrophic gastritis (CAG) (P = 0.001).

CONCLUSIONSThe results of this population-based study suggest that the virulence factors of Hp may be related to the development of GC in a Chinese high-risk population. The recomLine analysis may serve as a tool for identification of Hp strains and prediction of high-risk population of GC.

Adult ; Antibodies, Bacterial ; blood ; Female ; Gastritis ; blood ; immunology ; microbiology ; Gastritis, Atrophic ; blood ; immunology ; microbiology ; Helicobacter Infections ; blood ; immunology ; Helicobacter pylori ; Humans ; Male ; Middle Aged ; Precancerous Conditions ; blood ; immunology ; microbiology ; Stomach Neoplasms ; blood ; immunology ; microbiology

OBJECTIVETo investigate the associations of serum gastrin-17 (G-17) concentration with helicobacter pylori (Hp) infection.

METHODSA (13)C-urea breath and ELISA test to determine the Helicobacter pylori status and to detect the serum gastrin concentration was conducted in 242 villagers in Linqu of Shandong Province, a high gastric cancer prevalence area in China.

RESULTSOf 242 subjects, 65 of 111 were found Hp-positive in males (58.56%), compared with 65 of 131 in females (49.62%) (chi(2) = 1.932, P = 0.165). The statistical difference was not observed among different age groups (chi(2) = 4.185, P = 0.123). The average level of G-17 among 242 subjects was (24.43 +/- 25.46) pmol/L and it was statistically higher in females (29.87 +/- 28.18) pmol/L than that in males (18.01 +/- 20.11) pmol/L (Z = -3.618, P < 0.001). However, there was no statistical difference found among age groups (chi(2) = 1.948, P = 0.378). The G-17 level in Hp-negative group (35.50 +/- 30.92) pmol/L was observed significantly higher than in Hp-positive group (14.90 +/- 13.79) pmol/L (Z = 5.368, P = 0.0001).

CONCLUSIONThe G-17 concentration was found higher in Hp-negative subjects than in Hp-positive subjects, and higher in female than in male, but no difference was found among age groups.

Adult ; China ; epidemiology ; Female ; Gastrins ; blood ; Helicobacter Infections ; blood ; epidemiology ; Helicobacter pylori ; isolation & purification ; Humans ; Male ; Middle Aged ; Rural Population ; Sampling Studies

OBJECTIVETo repair segmental mandibular defects with autogenous bone marrow stromal cells (BMSCs) and coralline hydroxyapatite.

METHODSIsolated BMSCs were in vitro expanded and osteogenically induced. In 11 canines, a 3 cm segmental mandibular defect in right mandible was created. Five canine's defects were repaired with cell-scaffold constructs made from induced BMSCs and coralline hydroxyapatite (CHA); Others were repaired with CHA as control. The engineered bone was evaluated by X-ray, CT, gross and histological examination, biomechanical test 12, 26, 32 weeks post-operation respectively.

RESULTSBMSCs grew well on the CHA. X-ray and CT images showed better callus formation at connection sites in experimental group over time while worse formation at connection sites eventually in control group. At 32 weeks post-operation in experimental group, the defects were well repaired grossly. Histologically, there were bony healing and lamellar bone formation, in experimental group fibrous healing and woven bone formation in control group. Biomechanical test revealed no significant difference between experimental group and normal control group.

CONCLUSIONSCanine segmental mandibular defects can be ultimately repaired with the tissue-engineered bone generated by autogenous osteogenic BMSCs and CHA scaffold.

Animals ; Bone Marrow Cells ; cytology ; Bone Substitutes ; Ceramics ; therapeutic use ; Dogs ; Hydroxyapatites ; therapeutic use ; Mandible ; physiology ; Mandibular Injuries ; pathology ; surgery ; Mesenchymal Stromal Cells ; cytology ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo evaluate the relationship between cyclooxygenase-2 (COX-2) methylation and expression, and precancerous gastric lesions.

METHODSMethylation status of COX-2 was evaluated by quantitative denaturing high performance liquid chromatography (DHPLC) in 1201 subjects with different gastric lesions. COX-2 expression was assessed by immunohistochemistry and Helicobacter pylori (H pylori) infection status was determined by 13C-urea breath test (13 C-UBT).

RESULTSThe percent of COX-2 methylation was increased steadily with the severity of gastric lesions, showing 10.6% of which with superficial gastritis/chronic atrophic gastritis (SG/CAG), 11.8% with intestinal metaplasia (IM) and 13.8% with indefinite dysplasia/dysplasia (Ind DYS/DYS) (chi2 = 8.312, P = 0.016). Stratified analysis indicated that the percents of COX-2 methylation in subjects with H pylori negative still increased with the severity of gastric lesions,of 8.8% in SG/CAG, 10.6% in IM and 14.1% in Ind DYS/DYS (chi2 = 6.629, P= 0.036). Moreover,the methylated proportion of COX-2 was negatively associated with the expression in gastric lesions, from 13.3% with mild expression to 7.6% with strong expression (chi2 = 10.400, P = 0.015).

CONCLUSIONOur findings indicated that COX-2 methylation was significantly associated with precancerous gastric lesions and H pylori infection, suggesting that promoter methylation of COX-2 might play an important role in the progression of gastric lesions.

Adult ; China ; epidemiology ; Cyclooxygenase 2 ; genetics ; metabolism ; DNA Methylation ; Female ; Gastric Mucosa ; metabolism ; pathology ; Helicobacter Infections ; epidemiology ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Promoter Regions, Genetic ; Stomach Diseases ; epidemiology ; metabolism ; pathology